Food and Fermentation Industries >

2021 , Vol. 47 >Issue 23: 46 - 51

DOI: https://doi.org/10.13995/j.cnki.11-1802/ts.027387

Purification and application of a transglutaminase mutant

  • SONG Xiaoping ,
  • SHENG Hefang ,
  • YU Yin ,
  • MA Ruotong ,
  • PAN Lina
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  • 1(Department of Pharmacy,Anhui Medical College,Hefei 230061,China)
    2(Anhui Engineering Research Center of Recombinant Protein Pharmaceutical Biotechnology,Hefei 230022,China)

Received date: 2021-03-16

  Revised date: 2021-04-14

  Online published: 2021-12-31

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Abstract

To study the purification, glycosylation and catalytic application of recombinant Microbial Transglutaminase Mutant (MTG-MUT2), the high-density fermentation of Pichia pastoris GS115 was used for the expression of the MTG-MUT2. Firstly, the crude enzyme was obtained by high density fermentation of Pichia pastoris, and the MTG-MUT2 was purified by histidine labeled affinity chromatography with the enzyme activity of 12.3 U/mL. Compared with the crude enzyme, the protein was purified 2.7 times with a recovery of 87.4%. Then, three different components of MTG-MUT2 were separated by molecular sieve, and the effect of glycosylation on enzyme activity was studied. It was found that the enzyme activity of MTG-MUT2 was mainly determined by its glycosylated protein, and the glycosylation of N282 site played a significant role in the enzyme activity. Finally, the MTG-MUT2 was applied for catalytic reaction of L-lysine (660 mmol/L) and L-glutamine (600 mmol/L). The characteristic peak of Raman scattering spectrum for the reaction product (glutamine dipeptide) catalyzed by MTG-MUT2 was 20% higher than that of the wild enzyme. These results indicate that the MTG-MUT2 has higher thermal stability and catalytic efficiency than the wild enzyme.

Key words： transglutaminase; L-lysine-L-glutamine; expression conditions; catalysis conditions; surface-enhanced Raman scattering; glycosylation

Cite this article

SONG Xiaoping , SHENG Hefang , YU Yin , MA Ruotong , PAN Lina . Purification and application of a transglutaminase mutant[J]. Food and Fermentation Industries, 2021 , 47(23) : 46 -51 . DOI: 10.13995/j.cnki.11-1802/ts.027387

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